Teaching molecular genetics using Paramecium and RNA interference: research-based learning and project ownership

Area of Science:

• Molecular Biology
• Genetics
• Cell Biology

Background:

• Research-based course design enhances student engagement and learning outcomes.
• Paramecium, a single-celled eukaryote, is a valuable model organism for classroom research.
• RNA interference (RNAi) is a powerful technique for gene silencing in various model organisms.

Purpose of the Study:

• To implement a research-centered project using RNA interference (RNAi) in Paramecium.
• To guide students in designing and creating an RNAi plasmid for targeted gene product depletion.
• To investigate the effects of gene depletion on cell morphology and swimming behavior in Paramecium.

Main Methods:

• Students designed and constructed RNAi plasmids targeting specific genes in Paramecium.
• Techniques included database searches, primer design, plasmid generation, subcloning, and bacterial screening.
• Control and gene-depleted Paramecium cells were analyzed for changes in morphology, swimming behavior, and RNA transcript levels.

Main Results:

• Successful depletion of targeted gene products (IFT38/40, IFT140, KATNIP) was achieved in Paramecium.
• Depleted cells exhibited significantly slower swimming speeds compared to control cells.
• No noticeable changes in cell morphology were observed in the depleted Paramecium.

Conclusions:

• Student-driven research projects using RNAi in Paramecium foster engagement and collaboration.
• This approach effectively teaches cutting-edge molecular techniques and generates publishable data.
• Paramecium serves as an effective model for advanced genetics studies, demonstrating the utility of RNAi.